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ANXA1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 3 and 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.

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Images

Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (AB261768), expandable thumbnail
  • Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (AB261768), expandable thumbnail
  • Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (AB261768), expandable thumbnail

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

Knockout validation

Sanger Sequencing, Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 3 and 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3

Recommended products

ANXA1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 3 and 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.

Key facts

Cell type

HeLa

Form

Liquid

Mutation description

Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 3 and 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3

Disease

Adenocarcinoma

Concentration
Loading...

Properties

Gene name

ANXA1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Cell culture

Biosafety level

EU: 2 US: 2

Viability

~ 80%

Adherent/suspension

Adherent

Gender

Female

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines

  • All seeding densities should be based on cell counts gained by established methods.

  • A guide seeding density of 2x104 cells/cm2 is recommended.

  • Cells should be passaged when they have achieved 80-90% confluence.

Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Notes

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Culture medium:  DMEM (High Glucose) + 10% FBS  

Initial handling guidelines:

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.

2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.

3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.

4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.

5. Once confluent passage into an appropriate flask at a density of 2x10E4 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.  

Subculture guidelines:

• All seeding densities should be based on cell counts gained by established methods.

• A guide seeding density of 2x10E4 cells/cm2 is recommended.

• Cells should be passaged when they have achieved 80-90% confluence.

• Do not allow the cell density to exceed 7x10E4 cells/cm2.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Annexin A1 also known as ANXA1 or annexin 1 is a multifunctional protein with a molecular mass of approximately 37 kDa. This protein belongs to the annexin family and is distributed widely in various tissue types including the central nervous system respiratory system and immune cells. Annexin A1 functions by binding to phospholipids in a calcium-dependent manner influencing membrane-related processes and signal transduction. Its expression can be examined using techniques like annexin A1 immunohistochemistry and annexin A1 ELISA to study protein localization and concentration.

Biological function summary

Annexin A1 is involved in modulating inflammatory responses and cell proliferation. It acts as a mediator in the resolution of inflammation by promoting apoptotic cell clearance and regulating leukocyte migration. Annexin A1 participates in complex interactions often associating with cell membrane phospholipids to exert its effects. It contributes significantly to cell adhesion migration and intracellular signaling events which are important for maintaining homeostasis and proper immune function.

Pathways

Annexin A1 plays a significant role in the glucocorticoid signaling pathway and the formation of the cytoskeleton. It interacts with proteins like lipocortin 1 and phospholipase A2 impacting processes such as arachidonic acid metabolism and cytoskeletal reorganization. These interactions underline annexin A1's involvement in regulating inflammation and maintaining cellular architecture linking it to cellular processes that govern stress responses and cell movement.

Associated diseases and disorders

Annexin A1 exhibits relevance to inflammatory diseases and cancer. It has been implicated in conditions such as asthma where it modulates inflammatory cell recruitment and activation. In cancer especially in breast cancer annexin A1 influences tumor progression by interacting with proteins like COX-2 affecting tumor cell invasion and metastasis. The protein's role in these conditions highlights its potential as both a biomarker and therapeutic target helping in understanding disease mechanisms and developing treatment strategies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (ab261768), expandable thumbnail

    Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (ab261768)

    Allele-1: 16 bp deletion in exon 3.

  • Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (ab261768), expandable thumbnail

    Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (ab261768)

    Allele-2: 1 bp insertion in exon 3.

  • Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (ab261768), expandable thumbnail

    Sanger Sequencing - Human ANXA1 (Annexin A1) knockout HeLa cell line (ab261768)

    Allele-3: Insertion of the selection cassette in exon 3.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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